Immunofluorescence analysis of Phospho-c-Kit / CD117 pTyr936 Antibody was done on 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Phospho-c-Kit / CD117 pTyr936 Antibody (44500G) at 1µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa flour 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing cytoplasmic and nuclear localization. Panel e is a no primary antibody control. The images were captured at 40X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of human c-Kit that contains tyrosine 936. The sequence is conserved in cow.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/million cells|
|Western Blot (WB)||2-3 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
c-Kit, also known as CD117 and stem cell factor receptor, is a 145 kDa transmembrane tyrosine kinase encoded by the c-Kit proto-oncogene. c-Kit acts to regulate a variety of biological responses including cell proliferation, apoptosis, chemotaxis and adhesion. Ligand binding to the extracellular domain leads to autophosphorylation on several tyrosine residues within the cytoplasmic domain, and activation. c-Kit mutations correlate with tumor growth and progression in a variety of cancers including mast cell disease, gastrointestinal stromal tumor, acute myeloid leukemia, Ewing sarcoma, and lung cancer. Phosphorylation at tyrosine 703 of c-Kit allows binding of Grb2 and activation of the Ras-Raf-ERK1 and 2 signaling pathway.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
CADM1 controls actin cytoskeleton assembly and regulates extracellular matrix adhesion in human mast cells.
44-500G was used in western blot to study the role of CADM1 in cytoskeleton assembly and regulates extracellular matrix adhesion in human mast cells
|Moiseeva EP,Straatman KR,Leyland ML,Bradding P||PloS one (9:null)||2014|